Thermo Scientific Richard-Allan Scientific Wright-Giemsa Stain Solution Instructions for Use

For in vitro diagnostic use. For differential staining of blood smears and bone marrow specimens.

Specimen Collection Results Fresh blood film or fresh EDTA anticoagulated blood film and bone marrow films. Specimens should be air Erythrocytes – Pale Pink dried after smear has been prepared and fixed in absolute methanol for 15 seconds. Eosinophilic Granules – Reddish Orange Leukocyte Nuclei – Purple Mode of Action Cytoplasm – Bluish Purple Neutrophilic Granules – Light Purple The Thermo Scientific™ Richard-Allan Scientific™ Wright-Giemsa Stain Solution is a mixture of several thiazin dyes in a methanol solvent. Ionic and nonionic forces are involved in the binding of these dyes. The staining Discussion solution has anionic and cationic properties. The negatively charged phosphoric acid groups of DNA attract the purple polychromatic cationic dyes to the nuclei. The blue basophilic granules are stained by the polychromatic Wright-Giemsa Stain Solution should be stored at room temperature. This staining reagent is for “In Vitro” use cationic dyes. Cationic cellular components, such as erythrocytes and eosinophilic granules, are stained by the only. Refer to the Safety Data Sheet for Health and Safety Information. All reagents are stable and should not form red and pink anionic dyes. The buffers used in the staining procedure liberate and activate dye ions allowing precipitants under ordinary storage parameters. It is recommended that the Wright-Giemsa Stain Solution be them to chemically bond with specific cellular components. When staining blood and bone marrow smears, the discarded after each use. All dyes used in this formulation have been certified by the Biological Stain Commission. pH of the staining solution and/or buffer is a critical factor. Technical Comments Technical Procedure Thicker films and bone marrow preparations will require longer staining times. Because there is variation in the Immersion Staining Protocol pH of tap water, use of tap water in the procedure may hinder staining results. Distilled or deionized water should 1. Thoroughly dry blood or bone marrow smears. be used during the staining procedure. The “ripening” of the polychromed dye is a continuous chemical reaction. Therefore, the stock solution should not be used or diluted after the expiration date. The staining procedure may 2. Fix smears in absolute methanol for 15 seconds to 5 minutes require modification to suit personal preference. Blood films which have not been thoroughly air dried before 3. Stain smears in Wright-Giemsa Stain Solution for 1 minute. staining may show sloughing of cells from slide. 4. Rinse smears in Phosphate Buffer pH 6.6 solution (Catalog #89032) for 5 minutes – do not agitate slide. References 5. Rinse smears briefly in running deionized water (5-6 dips). 1. Raphael, S.S. Lynch’s Medical Laboratory Technology, Fourth Edition. Saunders Company, Philadelphia, PA, 6. Air dry smears. 1983. 7. Examine smears under a microscope. 2. Lillie, R.D. H.J. Conn’s Biological Stains, Ninth Edition. Williams and Watkins Company, Baltimore, MD, 1977. 3. Brown, B.A. Hematology: Principles and Procedures, Fourth Edition. Lea and Febiger Company, Philadelphia, Horizontal Staining Protocol PA, 1984 1. Place slide with thoroughly dried film on a horizontal staining rack. Ordering Information 2. Flood smear with absolute methanol for 15-30 seconds and then drain. 3. Flood smear with Wright-Giemsa Stain Solution for 45 seconds. Product Size Qty. REF 4. Add equal amount of Phosphate Buffer pH 6.6 solution to smear and let stand for 1 minute. Wright-Giemsa Stain 32 oz 1 89013 5. Rinse smear thoroughly with Phosphate Buffer pH 6.6 solution. Wright-Giemsa Stain 1 gal 1 89014 6. Air dry and examine under a microscope. Wright-Giemsa Stain 2.5 gal 1 89017 Phosphate Buffer pH 6.6 (powder) 9.25 g 12/cs 89032

Anatomical Pathology Richard-Allan Scientific Thermo Shandon Limited Subsidiary of Thermo Fisher Scientific Subsidiary of Thermo Fisher Scientific 4481 Campus Drive, Kalamazoo, MI 49008 Tudor Road, Manor Park, Phone: 1 (800) 522-7270 Runcorn, Cheshire WA7 1TA, UK Phone: + 1 (269) 544-5600 Phone: +44 (0) 1928 534 050 IS89013 R10/15 www.thermoscientific.com/pathology Fax: + 44 (0) 1928 534 049